Biomedical subjects

D A Godfrey

Publications and source records attributed to D A Godfrey.

At least 19 recordsLinked to original sources

PubMed1992

Sampling fluid from slice chambers by microsiphoning.

We have developed a simple technique that continuously samples perfusion fluid from an in vitro slice chamber and permits simultaneous electrophysiological recordings from a slice. This procedure uses a microsiphon that diverts approximately 1.5% of the total effluent of artificial cerebrospinal fluid from an interface slice chamber into a collection tube at a lower level outside the chamber. The tip of the microsiphon may be moved anywhere within the chamber without disturbing the slice. For optimum sampling of the release of substances from one area of a slice, the tip is positioned immediately downstream from the location being studied. Concentrations of several amino acids were measured to identify significant spatial and temporal characteristics of the technique and to demonstrate that spontaneous and induced release of amino acids from slices can be measured with adequate sensitivity and time resolution.

Amino Acids↗

PubMed1992

Connections between the cochlear nuclei in guinea pig.

This study provides a detailed analysis of the appearances and distributions of neurons projecting from one cochlear nucleus to the other. Injections of wheatgerm agglutinin conjugated to horseradish peroxidase were made into ventral or dorsal cochlear nucleus of the guinea pig. Retrogradely labeled cells in the opposite cochlear nucleus were examined and quantified. Three major categories of labeled cells were discerned on the basis of their soma shape: elongate, round-to-oval, and polygonal. All injections resulted in widespread labeling of cells in all of these categories, but especially round-to-oval cells, in the opposite ventral cochlear nucleus and sparse labeling in the dorsal cochlear nucleus. The results suggest that there is a significant cochlear nucleus commissural projection involving heterogeneous cell types which could have diverse functions in binaural auditory signal processing.

Animals↗

PubMed1991

Distributions of choline acetyltransferase and acetylcholinesterase activities in the retinal layers of the red-tailed hawk and road runner.

The activities of choline acetyltransferase and acetylcholinesterase were assayed in submicrogram samples from layers of red-tailed hawk and road runner retina. Both enzyme activities were concentrated in and near the inner plexiform layer. Within the inner plexiform layers of both species, activities of each enzyme were concentrated in two bands, one in each half of this layer. Little choline acetyltransferase activity was found superficial to the middle third of the inner nuclear layer. The distributions of acetylcholinesterase activities corresponded well to those of choline acetyltransferase, except in the outer plexiform layer and the outer margin of the inner nuclear layer of the hawk. These distributions of enzyme activities indicate that populations of amacrine cells in the retinae of these species are cholinergic. In addition to these same cells and presumably cholinoceptive amacrine and ganglion cells, acetylcholinesterase activity in the hawk was associated with a population of horizontal cells that may be unrelated to synaptic cholinergic neurotransmission. Choline acetyltransferase activities associated with amacrine somata and processes were about four times greater in the hawk than in the road runner, suggesting important differences in the density and function of cholinergic elements between species. Possible synaptic relationships in the inner plexiform layer consistent with the interspecies differences in enzyme activities are considered.

Acetylcholinesterase↗

PubMed1991

Descending projections to the dorsal and ventral divisions of the cochlear nucleus in guinea pig.

The origins of extrinsic projections to the guinea pig dorsal and ventral cochlear nuclei were identified by examining the retrograde transport of horseradish peroxidase conjugated to wheatgerm agglutinin following its injection into each of these divisions. Major projections originated in periolivary regions of the superior olivary complex, the contralateral cochlear nucleus and the inferior colliculus. There was no contribution from the nuclei of the lateral lemniscus to these pathways. The heaviest projection from the periolivary regions to both divisions of the cochlear nucleus arose bilaterally in the ventral nucleus of the trapezoid body. The ipsilateral lateral nucleus of the trapezoid body also projected heavily to dorsal and ventral cochlear nucleus. In addition, the ventral cochlear nucleus received a substantial projection from the dorsal aspect of the ipsilateral dorsomedial periolivary nucleus. Projections originating bilaterally in the central nucleus of the inferior colliculus terminated in the deep layers of dorsal cochlear nucleus. These projections appear to be more strongly ipsilateral and specific than those reported in the cat.

Animals↗

PubMed1990

Distributions of choline acetyltransferase and acetylcholinesterase activities in the retinal layers of pigeon red and yellow fields.

The activities of choline acetyltransferase and acetylcholinesterase were assayed in submicrogram samples from layers of pigeon retina. Red and yellow fields were sampled separately to investigate quantitatively the relationship between these enzymes of acetylcholine metabolism and the gradient of inner plexiform layer complexity, increasing from the yellow field to the red. Choline acetyltransferase and acetylcholinesterase activities were concentrated in and near the inner plexiform layer, within which two peaks of activity for each enzyme were obtained. The distributions of enzyme activities indicate that populations of amacrine cells in the pigeon retina are cholinergic. The quantitative similarities between the enzyme activities in red and yellow fields suggest that the cholinergic system may not be specifically involved in the increase in inner plexiform layer complexity across the pigeon retina.

Acetylcholinesterase↗

PubMed1990

Enzymes of transmitter and energy metabolism in rat middle ear and extraocular muscles.

To further investigate the peculiar characteristics of the middle ear and extraocular muscles, compared to the extensively studied skeletal muscles of the limbs, activities of enzymes of transmitter and energy metabolism were measured in homogenates of these muscles from albino and pigmented rats. These activities were compared to those for a masticatory muscle and for three hindlimb muscles chosen for their preponderance of either slow oxidative, fast glycolytic, or fast oxidative glycolytic fibers. Activities of the neuromuscular transmitter enzymes choline acetyltransferase and acetylcholinesterase were relatively very high in the extraocular and middle ear muscles. The activity of malate dehydrogenase, an enzyme of oxidative energy metabolism, was as high in the extraocular, masticatory and stapedius muscles as in the oxidative hindlimb muscles, but was lower in tensor tympani. The activity of lactate dehydrogenase, an enzyme of glycolytic energy metabolism, was remarkably low in both middle ear muscles. The results are consistent with high innervation density in the extraocular and middle ear muscles, and highly oxidative metabolism in the extraocular and stapedius muscles. Metabolic differences between the stapedius and tensor tympani suggest a relatively more active role for the former in the function of the rat middle ear.

Animals↗

PubMed1990

Contribution of centrifugal innervation to choline acetyltransferase activity in the cat cochlear nucleus.

Using a quantitative microchemical mapping approach combined with surgical cuts of fiber tracts, the contributions of centrifugal pathways to choline acetyltransferase activity were mapped three-dimensionally in the cat cochlear nucleus. Large reductions of choline acetyltransferase activity, averaging 70%, were measured in almost all parts of the lesion-side nucleus following transection of virtually all its centrifugal connections. More superficial cuts, penetrating just through the olivocochlear bundle, also led to significant reductions of enzyme activity, especially most rostrally in the anteroventral cochlear nucleus and superficial granular region, where the reductions were similar to those following the complete cuts. Lesions encroaching upon the superior olivary complex gave bilateral effects. Transverse cuts between rostral and caudal parts of the cochlear nucleus gave some small effects. The results suggest that, as in rats, most choline acetyltransferase activity in the cat cochlear nucleus is associated with its centrifugal innervation. However, unlike the situation in rats, the enzyme activity in cats is related more to olivocochlear branches than to ventral fibers in the trapezoid body region. Also, the choline acetyltransferase activity related to olivocochlear collateral innervation is much less uniformly distributed within the cochlear nucleus in cats than in rats.

Animals↗

PubMed1990

Quantitative distributions of aspartate aminotransferase and glutaminase activities in the guinea pig cochlea.

Distributions of aspartate aminotransferase and glutaminase activities in the guinea pig cochlea have been examined with use of quantitative microchemical techniques to evaluate their roles in cochlear energy metabolism and neurotransmission. Other enzyme activities analyzed were those of choline acetyltransferase and malate dehydrogenase. It is concluded that aspartate aminotransferase activity appears to be especially concerned with cochlear energy metabolism, while glutaminase activity may function in transmitter metabolism in the guinea pig cochlea. Neither enzyme shows a clear association with the olivocochlear bundle.

Animals↗

PubMed1990

Enzymes of transmitter and energy metabolism in cat middle ear muscles.

Activities of the enzymes choline acetyltransferase (ChAT) and acetylcholinesterase (AChE), which metabolize the neuromuscular transmitter acetylcholine, and malate and lactate dehydrogenase (MDH and LDH), enzymes of oxidative and glycolytic energy metabolism, respectively, were measured in the middle ear muscles of the cat. For comparison, the same enzyme activities were measured in extraocular muscle tissue and in three hindlimb muscles rich in either slow oxidative (soleus), fast glycolytic (white part of vastus lateralis), or fast oxidative glycolytic (plantaris) muscle fibers. ChAT and AChE activities were much higher in middle ear muscles than in hindlimb muscles, consistent with a denser neuromuscular innervation, as in extraocular muscles. By contrast, MDH and LDH activities were remarkably low in the middle ear muscles, lower than in any of the hindlimb muscles or the extraocular muscles. Denervation of the stapedius muscle by peripheral transection of the facial nerve resulted in decreases in all four enzyme activities without associated changes in the tensor tympani. Surgical ablation of the peripheral facial nerve supply to the stapedius muscle appears to be a feasible option for producing its denervation. The results suggest some rather specialized chemical characteristics for the middle ear muscles.

Acetylcholinesterase↗

PubMed1989

Quantitative distribution of six amino acids in rat retinal layers.

Concentrations of glutamate, aspartate, glutamine, glycine, GABA, and taurine were determined in samples microdissected from rat retinal layers and assayed by HPLC. Glutamate and glutamine were relatively high in the inner nuclear (INL) and ganglion cell (GCL) layers; aspartate was relatively high in the outer nuclear layer (ONL), outer plexiform layer, and INL. Distributions of glutamate and aspartate did not correlate well with those of enzymes involved in their metabolism. Glycine and GABA were highest in the inner plexiform layer, with increasing concentrations through the INL, and were relatively high in the GCL. Taurine was highest in the ONL.

Amino Acids↗

PubMed1989

Effect of septal ablation on choline acetyltransferase in the paraventricular nucleus.

To determine whether there is a cholinergic projection from the lateral septum to the paraventricular nucleus of the hypothalamus (PVH), a quantitative histochemical analysis of the effects of unilateral ablation (0.5 microliter of a 10 micrograms/ml solution of ibotenic acid) of the lateral septum (LSV) on the choline acetyltransferase (ChAT) activity of the PVH and surrounding perinuclear area was undertaken. Comparisons of the ChAT activity of the PVH region ipsilateral to the LSV lesion (lesioned side) with that of the PVH contralateral (nonlesioned side) to the ablated LSV showed a 20% reduction (p less than 0.05) of ChAT activity in 2 areas of the PVH on the lesioned side. When ChAT activity of LSV-ablated and intact control rats was compared, a 20% difference in activity (p less than 0.05) was noted in four areas on the nonlesioned side and a 35% difference (p less than 0.05) of ChAT activity was noted in five regions of the PVH on the lesioned side of LSV-ablated animals. Taken together these data suggest that the pathway from the LSV to the PVH contains a small, bilateral cholinergic component. However, the data also indicate that this is not the only cholinergic projection to the PVH.

Animals↗

PubMed1987

Quantitative inter-strain comparison of the distribution of choline acetyltransferase activity in the rat cochlear nucleus.

The distribution of choline acetyltransferase activity in the cochlear nucleus of Sprague-Dawley albino rats was quantitatively compared to those in two strains of pigmented rats, Long Evans hooded and Brown Norway, using microdissection and radiometric assay techniques. Although activities tended to be, on the whole, higher in the albino rats, the differences were fairly minor. The relative distributions of choline acetyltransferase activity were generally similar among the 3 rat strains, not only among regions, but also within regions. Stain for acetylcholinesterase activity in the cochlear nucleus also had a similar appearance among the 3 rat strains. These chemical results are consistent with previous anatomical and physiological studies suggesting that auditory differences between albino and pigmented animals may not be as great in the cochlear nucleus as in the superior olivary complex.

Animals↗

PubMed1987

Distributions of the activities of aspartate aminotransferase isoenzymes in rat retinal layers.

Distributions of the activities of the cytosolic (cAAT) and mitochondrial (mAAT) isoenzymes of aspartate aminotransferase were determined in rat retinal layers. cAAT was highest in the photoreceptor inner segments, inner nuclear layer and inner plexiform layer; mAAT was highest in the inner segments. The high activity in the inner segments indicates that both isoenzymes are involved in energy metabolism in addition to a possible role in neurotransmission.

Animals↗

PubMed1987

Distribution of activities of aspartate aminotransferase isoenzymes and malate dehydrogenase in guinea pig retinal layers.

Distributions of activity of the cytosolic (cAAT) and mitochondrial (mAAT) isoenzymes of aspartate aminotransferase and of malate dehydrogenase (MDH) were determined in guinea pig retinal layers. The distribution of total AAT activity (tAAT = cAAT + mAAT) and of mAAT activity correlated well (r = 0.88-0.91) with the distribution of MDH activity. mAAT activity was highest in the inner segments of the photoreceptors; there was a greater than twelve-fold difference between activity in that layer and in the inner retinal layers. cAAT activity was also highest in the inner segments, but the difference between the activity in the inner segments and the other layers was not nearly as great as with mAAT. cAAT activity was also relatively high in the outer nuclear layer, outer plexiform layer, and part of the inner plexiform layer. The high activity of cAAT, mAAT, and MDH in the inner segments indicates that all of these enzymes are involved in metabolic reactions related to energy production and/or to photoreceptive processes in the outer segments and, therefore, that the enzymes are probably involved in energy-related metabolism at synapses. However, other functions, including those related to neurotransmission, are not excluded.

Animals↗

PubMed1986

The distribution of choline acetyltransferase in the rat amygdaloid complex and adjacent cortical areas, as determined by quantitative micro-assay and immunohistochemistry.

The distribution of choline acetyltransferase (ChAT) within the amygdaloid complex has been studied to evaluate what should primarily represent the terminal field of the cholinergic projection from the basal forebrain. Two currently available methods have been combined for the comparison: immunohistochemistry with a monoclonal antibody against ChAT, by a double peroxidase-antiperoxidase procedure, and quantitative histochemistry involving micro-assay of the ChAT activity of contiguous microdissected samples. Both methods indicate prominent ChAT activity in the basolateral amygdaloid nucleus (especially rostrally), the nucleus of the lateral olfactory tract (especially layer II), and the amygdalohippocampal area. Regions of lower ChAT activity were not accurately represented by the immunohistochemistry, but could be discriminated by the quantitative assays. Lowest activity was found in the medial nucleus of the amygdala. Most other regions had activities at least as high as average brain or neocortex. Gradients of enzyme activity were found within several regions, including the basolateral and lateral amygdaloid nuclei and the nearby posterior piriform cortex. In the piriform cortex, a region of particularly high ChAT activity was found at its medial edge near the nucleus of the lateral olfactory tract. The immunohistochemical method shows a few intensely reactive somata in layer III within this zone. Comparison of the results seen with immunohistochemistry and quantitative histochemistry suggests an advantage in using them together, since their respective strengths and weaknesses tend to complement each other.

Amygdala↗

PubMed1986

Quantitative distributions of aspartate aminotransferase and glutaminase activities in the rat cochlea.

The intra-cochlear distributions of aspartate aminotransferase and glutaminase, prominent enzymes of aspartate and glutamate metabolism, have been studied by quantitative microchemical techniques. Also measured was choline acetyltransferase, the enzyme synthesizing acetylcholine, and a marker for the olivocochlear bundle. Aspartate aminotransferase activity was highest in the stria vascularis, about half this high in the organ of Corti synaptic (hair cell) zones, somewhat lower in the organ of Corti non-synaptic (Hensen's cell) zones, lower yet in Reissner's and lowest in the tectorial membrane. Glutaminase, on the other hand, had its highest activity in synaptic zones, about a third of that activity in the organ of Corti non-synaptic zones, and a barely detectable activity in Reissner's and tectorial membranes, and stria vascularis. Seven days after transection of the olivocochlear bundle, no significant difference was found between lesion- and control-side aspartate aminotransferase or glutaminase activities, even though no choline acetyltransferase activity remained in the lesion-side of the organ of Corti. Both the distribution of aspartate aminotransferase activity and the lesion results would seem to implicate it in energy more so than neurotransmitter metabolism. The distribution of glutaminase activity could be consistent with a role in neurotransmission; however, the lesion data were unable to demonstrate a specific association with the olivocochlear bundle.

Animals↗

PubMed1978

Distribution of gamma-aminobutyric acid, glycine, glutamate and aspartate in the cochlear nucleus of the rat.

The distributions of gamma-aminobutyric acid (GABA), glycine, glutamate and aspartate were measured in cochlear nuclei of two rats by quantitative histochemical mapping procedures. The levels and distributions in the two rats were comparable, and resembled those previously reported for cat cochlear nucleus. The results are consistent with a concept that these putative transmitter amino acids have similar levels and distributions in the cochlear nucleus among mammals.

Aminobutyrates↗

PubMed1977

Quantitative histochemical studies of the hypothalamus enzymes of acetylcholine metabolism.

The quantitative histochemical distribution of acetylcholinesterase and choline acetyltransferase activity has been measured in individual hypothalamic nuclei and median eminence, as well as in entire hypothalamic sections by a mapping technique. There was an 18-fold range of nuclear choline acetyltransferase activity with highest activities in the lateral preoptic nucleus and median eminence. There was a nine-fold range of nuclear acetylcholinesterase activity with highest activities in the lateral preoptic and magnocellular nuclei and lowest activity in the median eminence. The substantial gradients of choline acetyltransferase activity found in the hypothalamus indicate the importance of using a technique that provides an objective, permanent record of contiguous sample locations thereby allowing detailed analysis of tissue areas using, but not dependent on, anatomical boundaries.

Acetylcholinesterase↗